But it takes an hour and 47mins each time. The volts I cap at 10, Amps plateau out at around 100 (by themselves).

EC-150 is the machine that im hooking the semi dry apparatus up to to gain voltage. Its an ancient thing called thermo EC-150.

There is 20ul per lane, or do you mean ug?

Thanks so much, you seem to be very knowledgeable. I'm completely alone in my new lab with equipment Ive never used and no one to ask.

I think you should do some practice runs (unimportant sample/protein) and optimize. Semi-Dry is supposed to save you a lot of time versus the traditional tank-tranfer (1 hour to Overnight transfer times).

You should set constant amps at 0.5 or therabout (up to .7 should be fine); this plus short time protects the apparatus and proteins from excess heat.

Volts are probably too low, try 19-20V. Warning do this strictly with 20 minute time limit (go up to 25 max) and limit amps. 1 hour+ will burn the sandwich or proteins, even the apparatus (not huge danger, but a waste of your time/resources).

About the proteins 20ul doesn't tell me anything, ug/lane is much more informative. 20 ug and 20 minute transfer is more than enough. In my experience I load 80ug/lane (heterogenous mix of proteins) and 19 minutes at 17V is enough (0.5A max limit) to get 70-85% of the protein transfered (I stain the used gels with coomassie blue, imperial stain from Pierce actually).
With only large size proteins retained, all small proteins, less than 40 or 50kDa, transfer completely.

The main reason for the short time is that for transfer to occur, everything must stay wet with buffer, plus the convenience of saving yourself time. I don't think this will happen at 1 hour + transfer times.

If you can't limit the Amps, then perhaps you will have to keep volts down somewhat (like 12-14V and increase time 25-30 min max).

Yeah after a run that long I would expect the transfer buffer to be used up, I use very little simply saturating the buffer sandwich, but after the hour 47mins the sandwich is still quite saturated.

...... do you heat your samples at 95 or 100?

WOOW!!, hold it, my sandwiches are bone dry after semi-dry blotting 19-20min as described before. So that is your problem, this is not normal.

You probably have bad contact, perhaps the blot filter papers are not the right thickness, double/triple up and problem solved. Or maybe the little springs in the semi-dry blot apparatus are very compressed and not pushing the platinum anode out enough---just push them out for them to decompress, slowly and gently.

[Could also be buffer just isn't running buffer, no ions to run---check that the buffer makeup is actual running buffer recipe and concentration (proper water source was used).]

Clean the semi-dry apparatus (not the power source) with copious amount of DI water make sure buffer salts aren't beneath the plaforms (both lid and bottom), terminals (where power source wires make contact)---then leave them Upside Down over paper towels overnight to drip to complete dryness. Check for a springiness to the apparatus lid, the platinum coated platform should be bouncy, any flaky stuff and you have salt buffer build-up in there.

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Yes samples are boiled in Laemmli loading buffer for 5 minutes, then quickly centrifuged before loading on gel.

Remember after the blotting, you can coomassie or Imperial Blue stain the gel to check transfer efficiency.